Abstract Traumatic brain injury (TBI) is a substantial health care issue affecting both civilian and military populations. Although extremes of head injury are recognizable, mild to moderate TBI (MMTBI) is more difficult to diagnose leading to a need for the identification of circulating biomarkers of brain injury that may facilitate early identification. Using a rat model of TBI we identified a novel marker of neuronal injury, visinin like protein 1 (VILIP-1), that is released into blood rapidly after injury. To facilitate rapid measurement of circulating VILIP-1 we developed a combined lateral flow device (LFD) that provides detection of the biomarker in less than 20 min. Although initial testing in human TBI patients are promising there is a need for further testing in blood samples drawn clinically well characterized TBI patients soon after injury. The aim of the current proposal is to further validate the use of our LFD as a point of care diagnostic for TBI using a combination of archived serum samples from normal control subjects as well as patients with systemic traumatic injuries but no TBI and subjects with mild to moderate TBI and whole blood samples from patients being evaluated in the emergency department of Orlando Regional Medical Center. If successful, the proposed studies will validate the use of our VILIP-1 LFD for rapid in field detection of MMTBI and will provide the preliminary data to support a larger clinical trial of the device that could ultimately benefit both civilian and military populations.